Snf1p/Hxk2p/Mig1p pathway regulates hexose transporters transcript levels, affecting the exponential growth and mitochondrial respiration of Saccharomyces cerevisiae.

The Crabtree effect molecular regulation comprehension could help to improve ethanol production with biotechnological purposes and a better understanding of cancer etiology due to its similarity with the Warburg effect. Snf1p/Hxk2p/Mig1p pathway has been linked with the transcriptional regulation of the hexose transporters and phenotypes associated with the Crabtree effect. Nevertheless, direct evidence linking the genetic control of the hexose transporters with modulation of the Crabtree effect phenotypes by the Snf1p/Hxk2p/Mig1p pathway is still lacking. In this sense, we provide evidence that SNF1 and HXK2 genes deletion affects exponential growth, mitochondrial respiration, and transcript levels of hexose transporters in a glucose-dependent manner. The Vmax of the hexose transporters with the high transcript levels was correlated positively with the exponential growth and negatively with the mitochondrial respiration. HXT2 gene transcript levels were the most affected by the deletion of the SNF1/HXK2/MIG1 pathway. Deleting the orthologous genes SNF1 and HXK2 in Kluyveromyces marxianus (Crabtree negative yeast) has an opposite effect compared to Saccharomyces cerevisiae in growth and mitochondrial respiration. Overall, these results indicate that the SNF1/HXK2/MIG1 pathway regulates transcript levels of the hexose transporters, which shows an association with the exponential growth and mitochondrial respiration in a glucose-dependent manner.

Outcomes of pediatric living donor kidney transplantation: A single-center experience.

Renal transplantation is the treatment of choice for children with ESRD offering advantages of improved survival, growth potential, cognitive development, and quality of life. The aim of our study was to compare the outcomes of LDKT vs DDKT performed in children at a single center. Retrospective chart review of pediatric patients who underwent kidney transplantation from 2005 to 2014 was performed. Ninety-one renal transplants were accomplished, and 31 cases (38.27%) were LDKT, and in 96.7% of the cases, the graft was obtained through laparoscopy. Thirty-four receptors weighted <25 kg. LDKT group had statistically significant lower cold ischemia times than DDKT one. Complication rate was 9.67% for LDKT and 18.33% for DDKT. eGFR was better in LDKT. Patient survival rate was 100% for LDKT and 98.3% for DDKT, and graft survival rate was 96.7% for LDKT and 88.33%-80% for DDKT at a year and 5 years. Our program of pediatric kidney transplantation has achieved optimal patient and graft survival rates with low rate of complications. Living donor pediatric kidney transplants have higher patient and better graft survival rates than deceased donor kidney transplants.

Discovery, characterization and biological evaluation of a novel (R)-4,4-difluoropiperidine scaffold as dopamine receptor 4 (D4R) antagonists.

Herein, we report the synthesis and structure-activity relationship of a novel series of (R)-4,4-difluoropiperidine core scaffold as dopamine receptor 4 (D4) antagonists. A series of compounds from this scaffold are highly potent against the D4 receptor and selective against the other dopamine receptors. In addition, we were able to confirm the active isomer as the (R)-enantiomer via an X-ray crystal structure.

Synthesis and characterization of a series of chiral alkoxymethyl morpholine analogs as dopamine receptor 4 (D4R) antagonists.

Herein, we report the synthesis and structure-activity relationship of a series of chiral alkoxymethyl morpholine analogs. Our efforts have culminated in the identification of (S)-2-(((6-chloropyridin-2-yl)oxy)methyl)-4-((6-fluoro-1H-indol-3-yl)methyl)morpholine as a novel potent and selective dopamine D4 receptor antagonist with selectivity against the other dopamine receptors tested (<10% inhibition at 1µM against D1, D2L, D2S, D3, and D5).

Application of failure mode and effect analysis in laparoscopic colon surgery training.

AIM: To evaluate if application of failure mode and effect analysis (FMEA) to laparoscopy training can help surgeons acquire laparoscopy skills. METHODS: After preparing a FMEA matrix of laparoscopic sigmoidectomy, we have introduced it during three laparoscopy courses. Forty-eight surgeons, divided into 24 teams of two surgeons, have participated in three courses. During each course, every team has performed three laparoscopic sigmoidectomies in three experimental animals (1 OR session every day). Risk priority number (RPN) has been calculated for every surgery, and the results have been discussed at the end of each training day with all participants. RESULTS: We have observed a decline in the median RPN from 1339 during the first OR session through 62 during second OR session to reach 0 in the third OR session. Only two teams out of 24 were not able to reach a RPN of less than 300 during third OR session. When the type of failures were analysed, we have observed a shift from procedure-type failures to technical failures that depended on each participant technical abilities. CONCLUSION: Application of FMEA principles to laparoscopy training can help acquire non-technical skills necessary for safe laparoscopic surgery.

Comparison of single access devices during cut and suturing tasks on simulator.

BACKGROUND: Published comparisons of the different available laparoendoscopic single-site surgery (LESS) devices focused on its economic cost and technical aspects. With this study, we aimed to objectively compare the use of three different LESS access devices in controlled experimental tasks. MATERIALS AND METHODS: Twenty subjects participated in simulator trials. A cut and an intracorporeal suturing task were carried out. Three single access devices (SILS: SILS(TM) Port, GPN. GelPOINT Advanced Access Platform, and XCN: XCONE) were used according to a randomized nine-session schedule. Completion time was registered and performance objectively assessed with task-specific rating scales. Two blinded expert raters worked over video recordings of the hands-on sessions. RESULTS: Participants showed improvement with all devices on total cut completion times and significantly for SILS (P = 0.017). The GPN decreased its adapted Global Rating Scale score (P = 0.002) from the first (W1) to the last week (W9). On the suture task, XCN constituted the device with longer completion times compared with SILS (P < 0.001) and to GPN (P < 0.001). There was significant improvement in times from W1 to W9 with XCN (P < 0.001), SILS (P = 0.003), and GPN (P < 0.001). On average summative score, we observed significant improvement in performance with all devices from W1 to W9 (SILS: P = 0.003; GPN: P = 0.001; and XCN: P < 0.001). CONCLUSIONS: Although we advise surgeons to focus on the specific procedures and patient characteristics to select the most adequate access device to maintain procedural safety standards, single-use devices appear to confer an easier adaptation to LESS surgery.

Early postoperative complications of transvaginal access in minimally invasive sigmoid colon procedures.

OBJECTIVES: The aim of the study was to evaluate early (the first 30 days) postoperative complications after transvaginal resection of the sigmoid colon. MATERIAL AND METHODS: A total of 23 laparoscopy-assisted transvaginal resections of the sigmoid colon and 1 NOTES transvaginal sigmoid resection were performed in the course of 3 years. Postoperative complications were recorded in a prospective manner. RESULTS: In the group of 24 patients operated on using the transvaginal approach, 6 (25%) complications were recorded, including 3 urinary tract infections, 2 vaginal bleedings, and 1 abdominal trocar site hernia. CONCLUSION: Early postoperative complication rate after transvaginal resection of the sigmoid colon is relatively low and the clinical complications are not severe.

Ureteral obstruction swine model through laparoscopy and single port for training on laparoscopic pyeloplasty.

This study aims firstly to assess the most adequate surgical approach for the creation of an ureteropelvic juntion obstruction (UPJO) animal model, and secondly to validate this model for laparoscopic pyeloplasty training among urologists. Thirty six Large White pigs (28.29±5.48 Kg) were used. The left ureteropelvic junction was occluded by means of an endoclip. According to the surgical approach for model creation, pigs were randomized into: laparoscopic conventional surgery (LAP) or single port surgery (LSP). Each group was further divided into transperitoneal (+T) or retroperitoneal (+R) approach. Time needed for access, surgical field preparation, wound closure, and total surgical times were registered. Social behavior, tenderness to the touch and wound inflammation were evaluated in the early postoperative period. After ten days, all animals underwent an Anderson-Hynes pyeloplasty carried out by 9 urologists, who subsequently assessed the model by means of a subjective validation questionnaire. Total operative time was significantly greater in LSP+R (p=0.001). Tenderness to the touch was significantly increased in both retroperitoneal approaches, (p=0.0001). Surgeons rated the UPJO porcine model for training on laparoscopic pyeloplasty with high or very high scores, all above 4 on a 1-5 point Likert scale. Our UPJO animal model is useful for laparoscopic pyeloplasty training. The model created by retroperitoneal single port approach presented the best score in the subjective evaluation, whereas, as a whole, transabdominal laparoscopic approach was preferred.

[Training and research in urologic laparoscopic surgery. Design of training programs and experimental models of renal tumor]. / Formación e investigación en cirugía laparoscópica urológica. Diseño de programas formativos y modelos experimentales de tumor renal.

We present our experience with surgical training programs development for basic and advanced laparoscopic urological surgery. Both training programs consist of 21 and 28 hours respectively. Basic surgical programs start with general knowledge of ergonomics and instrumentation, there after, attendants acquire basic skills on physical simulator. Posteriorly, techniques on animal model are undertaken, always assisted by an expert. Advanced activities start with surgical tasks on physical simulator. Posteriorly, reconstructive urological surgical techniques are undertaken on animal model, focused on partial nephrectomy, and always assisted by an expert tutor. We present our results on exophytic renal tumour model creation based chromatic Alginate.

[New technologies in partial nephrectomy]. / Nuevas tecnologías en la nefrectomía parcial.

OBJECTIVES: We aim to analyse the role of new technologies in management of small renal cancer. METHODS: We perform a non-systematic review of the literature in Medline, Cochrane Database of Systematic Reviews between period 2000-2012, using following mesh terms: partial nephrectomy, renal ablative technologies, and renal cancer. RESULTS: We don't review in this article ablative technologies such as cryotherapy, radiofrequency, as they are the subject of others manuscripts within this monographic issue. We focus on high intensity ultrasounds (HIFU) microwaves therapy, radiosurgery, laser and water jet dissection. CONCLUSIONS: New technologies in partial nephrectomy are under constant and vertiginous evolution. Although efficacy has been demonstrated in short term and isolated studies, more studies, better designed, with bigger sample size and longer follow up are needed.

Totally transvaginal resection of the descending colon in an experimental model.

BACKGROUND: Natural orifice translumenal endoscopic surgery (NOTES), although in its embryonic phase, is currently experiencing important developments. The technique has been successfully applied for cholecystectomies and appendectomies. However, several doubts exist as to the technical limitations and feasibility of NOTES in other clinical settings. METHODS: The authors have performed totally transvaginal colon resections in a sheep model. Although completion of the surgery was possible through the transvaginal route, the addition of a transumbilical laparoscope was used as an added safety measure. RESULTS: Totally transvaginal resection of the sigmoid colon was performed for two sheep with no intra- or postoperative complications. CONCLUSION: Totally transvaginal resection of the colon (pure NOTES) is feasible in a sheep model.

[Ergonomics in laparoscopic surgery and its importance in surgical training]. / Ergonomía en cirugía laparoscópica y su importancia en la formación quirúrgica.

Despite the many advantages that laparoscopic surgery has for patients, it involves a series of risks for the surgeon. These are related to the reduced freedom of movement and forced postures which lead to greater muscle fatigue than with conventional surgery. In laparoscopic surgery there are few references on the introduction of training programs in ergonomics, despite the numerous advantages demonstrated in other disciplines. The application of ergonomic criteria in the surgical field could have great benefits, both for surgeons and patients. In this work we attempt to review the existing literature and our experience to provide the surgeon with some ergonomic guidelines for body stance and positioning of equipment. We also present a training model based on ergonomics which we have introduced into the training activities carried out in our Centre.

Association of ABC and RENAL Scoring Systems With Morbidity in Partial Nephrectomy.

BACKGROUND/AIM: Different nephrometry scoring systems (NSSs) are used to evaluate the surgical complexity and outcomes of partial nephrectomy (PN) in patients with small renal tumors. This study aimed to assess the validity of nephrometry scoring systems towards aiding the preoperative planning of laparoscopic partial nephrectomy (PN). PATIENTS AND METHODS: Data of 77 patients who underwent partial nephrectomy at the Puerto Real University Hospital between January 2011 and December 2017 were retrospectively analyzed. Statistical analyses were carried out to determine whether there was an association between the complexity of the surgical procedure and the assigned nephrometry scores. RESULTS: Operative complications (bleeding volume, conversion to open surgery, perioperative bleeding, and postoperative fistula) were significantly associated with independent variables (age, sex, body mass index, radiological tumor size, and operative ischemia time) and with the classification of patients using arterial-based complexity (ABC) and radius endophytic/exophytic nearness anterior-posterior location (RENAL) scores. There was also a strong correlation between the RENAL and ABC scores [Cramer's V coefficient (0.682) and Fisher's test (p<0.0001)]. CONCLUSION: The RENAL and ABC scores are associated with the risk of the complexity of partial nephrectomy for T1 renal tumors, even for ≥T1b tumors and/or with complex anatomical features.

Effect of acclimatization on hemocyte functional characteristics of the Pacific oyster (Crassostrea gigas) and carpet shell clam (Ruditapes decussatus).

Most experimental procedures on molluscs are done after acclimatization of wild animals to lab conditions. Similarly, short-term acclimation is often unavoidable in a field survey when biological analysis cannot be done within the day of sample collection. However, acclimatization can affect the general physiological condition and particularly the immune cell responses of molluscs. Our aim was to study the changes in the hemocyte characteristics of the Pacific oyster Crassostrea gigas and the carpet shell clam Ruditapes decussatus acclimated 1 or 2 days under emersed conditions at 14 ± 1 °C and for 1, 2, 7, or 10 days to flowing seawater conditions (submerged) at 9 ± 1 °C, when compared to hemolymph withdrawn from organisms sampled in the field and immediately analyzed in the laboratory (unacclimated). The hemocyte characteristics assessed by flow cytometry were the total (THC) and differential hemocyte count, percentage of dead cells, phagocytosis, and reactive oxygen species (ROS) production. Dead hemocytes were lower in oysters acclimated both in emersed and submerged conditions (1%-5%) compared to those sampled in the field (7%). Compared to oysters, the percentage of dead hemocytes was lower in clams (0.4% vs. 1.1%) and showed a tendency to decrease during acclimatization in both emersed and submerged conditions. In comparison to organisms not acclimated, the phagocytosis of hemocytes decreased in both oysters and clams acclimated under submerged conditions, but was similar in those acclimated in emersed conditions. The ROS production remained stable in both oysters and clams acclimated in emersed conditions, whereas in submerged conditions ROS production did not change in both the hyalinocytes and granulocytes of oysters, but increased in clams. In oysters, the THC decreased when they were acclimated 1 and 2 days in submerged conditions and was mainly caused by a decrease in granulocytes, but the decrease in THC in oysters acclimated 2 days in emersed conditions was caused by a decrease in hyalinocytes and small agranular cells. In clams, the THC was significantly lower in comparison to those not acclimated, regardless of the conditions of the acclimatization. These findings demonstrate that hemocyte characteristics were differentially affected in both species by the tested conditions of acclimatization. The phagocytosis and ROS production in clams and phagocytosis in oysters were not different in those acclimated for 1 day under both conditions, i.e. emersed and submerged, and those sampled in the field (unacclimated). The THC was significantly affected by acclimatization conditions, so the differences between clams and oysters should be considered in studies where important concentrations of hemocytes are required. The difference in the immune response between both species could be related to their habitat (epifaunal vs. infaunal) and their ability of resilience to manipulation and adaptation to captivity. Our results suggest that functional characteristics of hemocytes should be analyzed in both oysters and clams during the first 1 or 2 days, preferably acclimated under emersed rather than submerged conditions.

Overcoming the recalcitrance for the conversion of kenaf pulp to glucose via microwave-assisted pre-treatment processes.

This study evaluates the pre-treatment of cellulose from kenaf plant to yield sugar precursors for the production of ethanol or butanol for use as biofuel additives. In order to convert the crystalline cellulosic form to the amorphous form that can undergo enzymatic hydrolysis of the glycosidic bond to yield sugars, kenaf pulp samples were subjected to two different pre-treatment processes. In the acid pre-treatment, the pulp samples were treated with 37.5% hydrochloric acid in the presence of FeCl(3) at 50 °C or 90 °C whereas in the alkaline method, the pulp samples were treated with 25% sodium hydroxide at room temperature and with 2% or 5% sodium hydroxide at 50 °C. Microwave-assisted NaOH-treatment of the cellulose was also investigated and demonstrated to be capable of producing high glucose yield without adverse environmental impact by circumventing the use of large amounts of concentrated acids i.e., 83-85% phosphoric acid employed in most digestion processes. The treated samples were digested with the cellulase enzyme from Trichoderma reesei. The amount of glucose produced was quantified using the Quantichrom(™) glucose bioassay for assessing the efficiency of glucose production for each of the treatment processes. The microwave-assisted alkaline pre-treatment processes conducted at 50 °C were found to be the most effective in the conversion of the crystalline cellulose to the amorphous form based on the significantly higher yields of sugar produced by enzymatic hydrolysis compared to the untreated sample.

Treatment of Metastatic Castration-resistant Prostate Cancer Patients With Abiraterone Acetate and Prednisone and Corresponding Survival Prognostic Factors.

BACKGROUND/AIM: The aim of the study was to evaluate the combined treatment with abiraterone acetate and prednisone (AA+P) in patients with castration-resistant prostate cancer (mCPRC), and to identify the survival prognostic factors. PATIENTS AND METHODS: Patients diagnosed with mCPRC not previously treated with chemotherapy and administered with AA+P were classified into two groups: those with lower and higher survival rates (at 30 months vs. 60 months). RESULTS: A total of 53 patients were studied at the time of mCRPC diagnosis. Patients with the highest survival rate had suffered prostate cancer for >45 months. At the time of initial prostate cancer diagnosis, they belonged to the risk groups 1-4, had pain intensity measured according to the brief pain inventory (BPI) scale of 0-2, were treated with AA+P>16 months, and had the following tumour marker serum levels: LDH baseline ≤163 U/l, alkaline phosphatase at 6 months ≤56 U/l and PSA at 6 months ≤0.95 ng/ml. CONCLUSION: Good response to treatment with AA+P for patients with mCRPC was demonstrated. Factors that contributed to the higher prognostic accuracy were time suffering from prostate cancer, the intensity of the pain measured by the BPI scale, the duration of AA+P treatment, and tumour marker levels.

[Validity studies in laparoscopic simulation. Methodology and design considerations.] / Validación en simulación laparoscópica. Consideraciones metodológicas y de diseño.

OBJECTIVES: To describe a roadmap of the most representative milestones and considerations in the validation of surgical simulators, especially those of laparoscopic surgery. And additionally, help determine when in this process a simulator can be considered as validated. METHODS: A non-systematic review was carried out searching terms like simulation, validation, training, assessment, skills and learning curve, as well as providing the experience accumulated by our center. RESULTS: An ideal classical validation process should consist of the following steps: fidelity, verification/calibration/ reliability, subjective and objective strategies. Baseline tests of fidelity and verification/calibration/ technological reliability are not always detailed in the simulation literature. A simulator can be considered validated if, at least, satisfactorily completed any of the two main objective strategies, that is, constructive and/or criterion validity. CONCLUSIONS: The methodologies to validate simulators as useful and reliable for the improvement of psychomotor/ technical skills are widely analyzed, although there is a variety of approaches depending on the scientific reference consulted, not being implemented equally in all works. This apparent arbitrariness should be considered in advance because it can lead the researcher to misunderstandings, especially when the simulator will be regarded as valid.

OBJETIVOS: Describir una hoja de ruta de los hitos y consideraciones más representativos en la validación de simuladores quirúrgicos, especialmente los de cirugía laparoscópica. Y adicionalmente contribuir a determinar en qué momento de este proceso puede considerarse un simulador como validado.MÉTODOS: Se realizó una revisión no sistemática con los términos simulación, validación, formación, entrenamiento, evaluación, habilidades y curva de aprendizaje, además de aportar la experiencia acumulada por nuestro centro. RESULTADOS: Un proceso ideal clásico de validación debería constar de los siguientes pasos: Fidelidad, Verificación/ Calibración/Fiabilidad, estrategias subjetivas y objetivas. Las pruebas de inicio tanto de Fidelidad como de Verificación/Calibración/Fiabilidad tecnológica no siempre están descritas de manera explícita en los trabajos de validación de simuladores. Un simulador puede considerarse validado si al menos ha completado satisfactoriamente una validación de cualquiera de los dos grandes bloques de tipo objetivo, es decir, constructiva y/o de criterio. CONCLUSIONES: Los métodos que permiten validar simuladores como útiles y fiables para la mejora de habilidades de tipo psicomotor/técnico están ampliamente documentados aunque existe cierta variedad de enfoques en función de la referencia científica que se consulte, no aplicándose por igual en todos los trabajos. Esta aparente arbitrariedad debería ser conocida de antemano porque puede llevar al investigador a ciertos equívocos, especialmente a la hora de afirmar cuándo el simulador se considera plenamente validado.

Anatomic study of human laryngeal ganglia: number and distribution.

We have studied 12 laryngeal nerves: six internal branches of the superior laryngeal nerve (ILN) and six recurrent laryngeal nerves (RLN) from three human adult larynges (two males and one female). After dissection of each individual laryngeal nerve using a surgical microscope, the nerves were preserved in 10% formalin, embedded in paraffin wax, serially sectioned transversely at a thickness of 10 microm and stained with hematoxylin and eosin. We found 2-4 ganglia associated with the ILN. At least two ganglia were always present (six out of six cases), the largest one being associated with the branch of the nerve innervating the vestibule and the smallest one associated with the branch innervating the aryepiglottic fold. Other ganglia were found associated with the branches for the glosso-epiglottic fold and vallecula (four out of six cases) and interarytenoid muscle (three out of six cases). The RLN showed from two to six ganglia, all of them located in its anterior terminal division. Two of the ganglia were located in the part of the nerve between the origin of the branches for the interarytenoid and lateral cricoarytenoid muscles (three out of six cases). The remaining ganglia were located close to or at the origin of the muscular branches innervating the intrinsic laryngeal muscles. The cytology of the ganglia reported suggests that they were all autonomic in nature, probably parasympathetic.

[Training in basic and advanced laparoscopy: Evaluation of a 30-year experience in a training program in Spain.] / Formación en cirugía laparoscópica urológica básica y avanzada: evaluación de 30 años de experiencia de un programa de formación en España.

The lack of globally established standards for learning urological laparoscopy has not prevented laparoscopic techniques from evolution and continuous development. Laparoscopy coexists with robotic surgery today, and in the last decade there have been many techniques that have undergone a boom with the use of a laparoscopic approach (total and partial nephrectomy, pyeloplasty, colposacropexy, etc.).We intend to evaluate the progressive incorporation of different surgical techniques in the laparoscopic learning program and, on the other hand, to analyze the evolution of training programs in urological laparoscopy to bring this type of techniques within the hospital surgical activity. We describe our 30-years experience in different training programs in urological laparoscopy that have been sponsored by the Spanish Association of Urology (AEU), and have undergone several validity studies to assess their capacity in order to evaluate effectively basic and advanced laparoscopic skills. We will also highlight the current and future trend towards training models based on surgical competences where individualized training, accreditation and specialization of tutors is crucial, and where the increase in the use of training and evaluation methods based on the simulation are increasingly common.